CN103224881B - Pre-treatment device and pre-treatment method for viable bacteria detection - Google Patents
Pre-treatment device and pre-treatment method for viable bacteria detection Download PDFInfo
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- CN103224881B CN103224881B CN201310102274.7A CN201310102274A CN103224881B CN 103224881 B CN103224881 B CN 103224881B CN 201310102274 A CN201310102274 A CN 201310102274A CN 103224881 B CN103224881 B CN 103224881B
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Abstract
The invention provides a pre-treatment device and a pre-treatment method for viable bacteria detection. The pre-treatment device comprises a shell, a lighting system, a refrigerating system and a sample module, wherein the lighting system, the refrigerating system and the sample module are arranged in the shell. The lighting system comprises a light source, a cooling fan and a radiator. The refrigerating system comprises semiconductor coolers, a cooling fan and a radiator. The sample module is arranged among the light source and the semiconductor coolers. Sample tube seats are arranged on the sample module and matched with sample tubes. Due to the sample tube seats, the sample tubes put into the sample tube seats are provided with tube wall faces facing toward the light source. The invention further provides a pre-treatment method for the viable bacteria detection and a detecting method for microorganisms. The purpose is that the pre-treatment device for samples to be detected is provided in prior to a PCR detection method and a Real time PCR detection method. Moreover, the pre-treatment method which is carried out through the pre-treatment device is easy and convenient to operate and good in effect.
Description
Technical field
The present invention relates to detection technique, particularly relate to the pretreatment process of a kind of living stems pretreatment unit and living stems.
Background technology
At present, in food safety, ambient water quality detection and prevention and control of diseases work, the detection technique of microorganism mainly contains conventional sense technology and molecular Biological Detection technology.Conventional sense technology is bacteria distribution cultural method mainly, and the positive rate of the method culturing bacterium is low, incubation time long (being generally 2-10 days), cannot meet current, high-throughput, responsive demand quick to microorganism detection, and molecular Biological Detection technology, as PCR and Real time round pcr mainly detects the DNA profiling of microorganism in sample, have fast, accurately, the advantage of sxemiquantitative or detection by quantitative, but what the method detected is copy for the DNA of microorganisms all in whole sample, comprise the DNA of the dead microbial release with bioactive DNA of bacteria and abiology and health significance, the result of detection and the actual deviation with the bacteria content of biologic activity can be caused like this, there is " false positive " result, have influence on and detect and assessment food, the accuracy of the bacteria content of biological sample in environmental sample and prevention and control of diseases.
At present, the dead DNA of bacteria in testing sample and DNA of bacteria of living cannot be distinguished for PCR and Real time PCR detection technique, before microorganism detection, first pre-treatment or pre-treatment are implemented to testing sample, the dead bacterium DNA in sample is degraded, to improve detection accuracy.That is, some have been had to report for the preprocessing process before PCR and Real time PCR detection.Such as: the open CN102605091A of patent, the denomination of invention EMA-PCR detection method of pathogenic bacteria viable bacteria " in plant's environment ", this detection method make use of and adds EMA reagent at testing sample, carry out pre-treatment under halogen light source illumination after, extract template DNA, carry out PCR detection with template DNA; The open CN102146480A of patent, denomination of invention " the living stems method of legionella and detection kit ", this detection method also utilizes EMA equally and to degrade in measuring samples DNA in dead mycetocyte, then extracts measuring samples DNA, by the viable count of legionella in fluorescence quantitative PCR detection sample.
Can see, comprise that above-mentioned what be disclosed in that interior prior art pays close attention to is all the operation of the preprocessing process of testing sample, and not open relevant pretreatment unit carries out specification pre-treatment accurately to testing sample.That is, the preprocessing process of prior art is all degraded first for testing sample by the DNA of EMA reagent to dead bacterium under the irradiation of light source, detects the DNA of the bacterium that lives after being separated.In operation, except selecting suitable treatment agent, under also needing that sample is placed in light source with means by rights, just can complete and reach expected result, the result that technician can only grope according to great many of experiments and accumulate, by experience, associated technical parameters and mode are set, be difficult to the Stability and veracity ensureing detected result.On the other hand, prior art needs to use incandescence-halogen lamp, and the power of this kind of light source is large, a large amount of heats can be produced, also the DNA of bacterium alive in testing sample can be caused to be degraded, to make subsequent detection be worth improper reduction, accurately can not reflect the legitimate reading of sample.Therefore, in DNA process in the dead bacterium of illumination degrading, usually need to use ice bath to carry out under cryogenic to control its degradation process, but this control device to low temperature can not control temperature accurately, and because of the error of operator, can melt completely under making ice bath be exposed to the irradiation of halogen lamp for a long time, thus the temperature of environment residing for testing sample is raised, cause sample fails, have a strong impact on the safety of testing sample.
Therefore, how to design a kind of pretreatment unit, can by technical parameters such as regulation and control light and temperature, realize standardization and the accuracy of the detected result of pretreatment process, be people's problem demanding prompt solutions always.
Summary of the invention
The object of the present invention is to provide a kind of pretreatment unit for living stems, pre-treatment can be carried out to the sample needing PCR and Real time PCR to detect, utilize effective control that this pretreatment unit can realize pretreatment parameter, and simple to operate, microorganism detection can be realized and standardize in advance and accuracy.
Present invention also offers a kind of pretreatment process utilizing described living stems pretreatment unit to carry out, this pretreatment process operating process is easy, and respond well.
The invention provides a kind of living stems pretreatment unit, this pretreatment unit comprises:
Housing;
Be arranged at the illumination system in this housing, refrigeration system and sample block;
Described illumination system comprises light source, radiator fan and scatterer;
Described refrigeration system comprises semiconductor cooler, radiator fan and scatterer;
Described sample block is arranged between described light source and semiconductor cooler;
Wherein, described sample block is provided with the sample tube seat matched with sample hose, and this sample tube seat can make inserted sample hose have towards the tube wall face of light source.
In pretreatment unit of the present invention, the top of described light source sets gradually scatterer and radiator fan composition illumination system, and described light source and described scatterer are formed and press laminating; Set gradually scatterer and radiator fan composition refrigeration system below described semiconductor cooler, and described scatterer and described semiconductor cooler form and press laminating.
In pretreatment unit of the present invention, when this pretreatment unit is vertically placed, described illumination system is arranged at the top in this pretreatment unit housing, described refrigeration system is arranged at the bottom in this pretreatment unit housing, the sample tube seat that described sample block is arranged can make the axis of inserted sample hose and horizontal direction be that 10-70 ° of overturning angle is placed, preferably 30 °, its angle of inclination, this angle can make sample hose fully accept the irradiation of light source, and make testing sample not easily excessive, do not pollute the mouth of pipe.
In pretreatment unit of the present invention, described sample block is provided with multiple sample tube seat, can be arranged in parallel between described multiple sample tube seat, sample tube seat also can be made to be the radial setting of disk.According to actual needs, can to arrange in sample block one or more groups sample tube seat, and the modes of emplacement often organized can make the blind end of sample hose towards unanimously, and the axis of sample hose and horizontal direction are 10-70 ° of angle places, also the blind end top of sample hose can be made staggered relatively to top, and the axis of sample hose and horizontal direction are 10-70 ° of angle places, do not do concrete restriction at this.
In pretreatment unit of the present invention, described sample block is provided with the illuminance transducer monitoring light illumination; And can the temperature sensor of monitoring system cooling system, and then regulate the work of semiconductor cooler, thus the temperature of detected sample in sample hose in more effective Quality control module, ensure the stability of DNA in testing sample.
In pretreatment unit of the present invention, described sample block is positioned in refrigeration system, formed by heat-transfer medium-heat conductive silica gel fat and described semiconductor cooler and press laminating, further, the upper surface of the scatterer of refrigeration system and this semiconductor cooler are formed by heat-transfer medium-heat conductive silica gel fat and press laminating.The heat of semiconductor cooler effectively can be passed to sample block by this heat-transfer medium-heat conductive silica gel fat, then passes to sample hose by sample block, and this heat conductive silica gel fat also can play the effect of tackiness agent simultaneously.
In pretreatment unit of the present invention, the housing of this pretreatment unit for having the tubular shell of upper shed, and can have the lid that can fasten this housing seal, and described illumination system is arranged on this lid; The housing of this pretreatment unit also can be provided with opening for sidewall, and be provided with can to the engaging support mechanism of this housing seal, and described sample block can take out through described opening or insert.
In pretreatment unit of the present invention, described sample block is arranged in inner walls by rail structure or between the radiator fan of refrigeration system of housing bottom and scatterer.Sample block easily along guide rail push-and-pull, can be drawn out or advances from pretreatment unit.
In pretreatment unit of the present invention, the distance between sample block and illumination system can carry out arranging adjustment as required, such as, and the illumination of light source, the character etc. that angle and testing sample are set of sample hose.In the specific embodiment of the present invention, the minimum perpendicular distance of described sample block and described light source can be made to be 2-10cm.
In pretreatment unit of the present invention, described light source is cold light source, such as: LED.Cold light source-LED, compared to incandescence-halogen lamp, has good optical property.
In pretreatment unit of the present invention, described housing is engineering plastics housing, and inner walls is mirror face stainless steel inwall.Being set to mirror face stainless steel material in the inner walls of this pretreatment unit can the light that sends of reflection source from different perspectives, avoids light loss, can irradiate sample hose fully, make its illuminating area enough large.
In pretreatment unit of the present invention, described sample block is aluminium alloy module.
In pretreatment unit of the present invention, this pretreatment unit also comprises control mainboard, the supply unit of controlled light system and refrigeration system, and the display unit of the illumination that can show in preprocessing process and/or state of temperature.
Present invention also offers a kind of pretreatment process of living stems, the pretreatment unit described in employing, the dead bacterium in testing sample is degraded, and described pretreatment process comprises the following steps:
The sample hose that testing sample and reagent are housed is placed in the sample tube seat that described pretreatment unit matches with sample hose;
Under light source, sequentially exposing irradiates sample hose, and Quality control module temperature is 2-8 DEG C, realizes the DNA of dead bacterium in degraded testing sample;
Wherein, described reagent comprises EMA or PMA.
Described reagent E MA is single nitrine Ethidium Bromide in the present invention, and PMA is nitrine bromination third ingot.
In pretreatment process of the present invention, according to kind and the character of itself of testing sample, can according to practical situation, select different temperature, intensity of illumination and light application time to control the DNA degradation of dead bacterium in testing sample, therefore not to repeat here.
Present invention also offers a kind of detection method of microorganism, comprise the following steps:
Pretreatment process of the present invention implements pre-treatment to testing sample, and the dead bacterium in testing sample is degraded;
Take out sample hose, DNA separation is carried out to this pretreated testing sample, and implements microorganism detection.
Compared with prior art, the present invention has following beneficial effect:
1, in prior art, can not accurately control the correlation parameter in the pretreatment process of testing sample, thus have a strong impact on the security of testing sample, and the defect of lack of standard in preprocessing process and inaccuracy, living stems pretreatment unit provided by the invention, pre-treatment can be carried out to the sample needing PCR and Real time PCR to detect, utilize effective control that this pretreatment unit can realize pretreatment parameter, and simple to operate, microorganism detection can be realized and standardize in advance and accuracy.
2, the pretreatment process that undertaken by the living stems pretreatment unit provided of the present invention, effectively can improve the accuracy that PCR, Real time PCR result detects, and operating process is easy, respond well.
Accompanying drawing explanation
Fig. 1 is the structural representation of the living stems pretreatment unit that the embodiment of the present invention provides.
Fig. 2 is the sectional side view of sample block in the living stems pretreatment unit that provides of the embodiment of the present invention.
Embodiment
Embodiment 1
Fig. 1 is the structural representation of the living stems pretreatment unit that the embodiment of the present invention provides, and Fig. 2 is the sectional side view of sample block in the living stems pretreatment unit that provides of the embodiment of the present invention.
The present embodiment discloses a kind of living stems pretreatment unit, concrete structure as depicted in figs. 1 and 2, this pretreatment unit comprise be made up of engineering plastics rectangle housing (inner walls is mirror face stainless steel material), the illumination system be arranged in housing, refrigeration system, aluminum alloy sample module 6; Described illumination system comprises LED-lamp 3, radiator fan 1 and scatterer 2; Described refrigeration system comprises semiconductor cooler 5, radiator fan 10 and scatterer 11; Described aluminum alloy sample module 6 is arranged between described LED-lamp 3 and semiconductor cooler 5.
Described illumination system is arranged at the top in this pretreatment unit housing, and described refrigeration system is arranged at the bottom in this pretreatment unit housing, and described aluminum alloy sample module 6 is arranged between illumination system and refrigeration system.Described aluminum alloy sample module 6 is provided with the sample tube seat matched with sample hose, sample tube seat is divided into four groups, often organize 6, wherein, in four groups of sample tube seats, every two groups of sample tube seats are towards consistent placement, wherein correspondingly with other its tube seat top of two groups of sample tube seats to place for two groups, and make its sample tube seat be that 30 degree of overturning angles are placed for making the axis of inserted sample hose and horizontal direction; The housing upper of described pretreatment unit is provided with upper cover, and sidewall is provided with opening, and be provided with can to the engaging support mechanism of this housing seal, and the described opening that described aluminum alloy sample module 6 can be provided with through sidewall takes out or inserts; Described aluminum alloy sample module 6 is arranged in inner walls by rail structure, and aluminum alloy sample module 6 described in it easily along guide rail 7 push-and-pull, can be drawn out or advance from pretreatment unit.The supply unit 9 of described control mainboard, controlled light system and refrigeration system, and the display unit 13 of the illumination that can show in preprocessing process and/or state of temperature is located at this pretreatment unit all outward; Minimum perpendicular distance between described aluminum alloy sample module 6 and described LED-lamp 3 is 8cm, and as shown in Figure 1, this minimum perpendicular distance is the vertical range of light source led-lamp 3 and illuminance transducer 4.
The top of described LED 3 is disposed with scatterer 2 and radiator fan 1, and described LED 3 and described scatterer 2 are formed and press laminating; Set gradually scatterer 11 and radiator fan 10 below described semiconductor cooler 5, and described scatterer 11 and described semiconductor cooler 5 press laminating.
Described aluminum alloy sample module 6 is provided with the illuminance transducer 4 monitoring light illumination, and the temperature sensor 12 of energy monitoring system cooling system.
Be positioned on the semiconductor cooler 5 of refrigeration system in described aluminum alloy sample module 6, formed by heat-transfer medium-heat conductive silica gel fat 8 and semiconductor cooler 5 and press laminating, further, the upper surface of the scatterer 11 of refrigeration system and this semiconductor cooler 5 are formed by heat-transfer medium-heat conductive silica gel fat 8 and press laminating.
Adopt the present embodiment living stems pretreatment unit to be example, the using method of living stems pretreatment unit of the present invention be described:
When using living stems pretreatment unit, the described opening that aluminum alloy sample module 6 is provided with by housing sidewall is taken out, after the sample hose that testing sample and reagent (EMA or PMA) are housed is placed in the sample tube seat coordinated with it, again aluminum alloy sample module 6 is advanced in pretreatment unit by the rail structure be arranged in inner walls, and by being provided with the illuminance transducer 4 monitoring light illumination in described aluminum alloy sample module 6, and the temperature sensor 12 of energy monitoring system cooling system regulates and controls intensity of illumination and regulates refrigeration system to control the temperature of aluminum alloy sample module 6, thus realize utilizing described pretreatment unit to carry out pre-treatment to testing sample.
Embodiment 2
Present embodiments provide and to adopt in embodiment 1 living stems pretreatment unit to the pretreatment process of legionella pneumophilia.
1, the selection of sample
Utilize buffering carbon powder yeast basic substratum to cultivate legionella pneumophilia (Lp1ATCC33152), preparing concentration is 1 × 10
8bacterium liquid to be measured.
2, the process of sample and DNA extraction
1) EMA of 5mg is prepared into the mother liquor of 100 μ g/mL;
2) in 1.5mL sample hose, add 450 μ L concentration is 1 × 10
8bacterium liquid to be measured, then to the EMA mother liquor adding 50 μ L, to sample hose, the final concentration of EMA is 10 μ g/mL;
3) above-mentioned sample hose is placed in the sample tube seat that living stems pretreatment unit matches with sample hose, the temperature of Quality control module is 4 DEG C, and uses LED exposure irradiation sample hose 5min, realizes the DNA of dead bacterium in degraded bacterium liquid to be measured.
Embodiment 3
Present embodiments provide the microorganism detection adopting the living stems pretreatment process of embodiment 2 to carry out pretreated bacterium liquid to be measured.
By pretreated for embodiment 2 bacterium liquid to be measured, carry out DNA separation with the centrifugal 15min of the rotating speed of 13000rpm, and utilize EMA-fluorescence quantifying PCR method to carry out living stems to legionella pneumophilia.
Comparative example
Adopt EMA-fluorescence quantitative PCR method to pretreated bacterium liquid to be measured respectively, slat chain conveyor counting process carries out microorganism detection to bacterium liquid to be measured.
1, in bacterium liquid to be measured, getting 100 μ L coats on substratum, is placed in 36 ± 1 DEG C, CO
2incubator cultivates 10 days, and counts the colony number on flat board.
2, get bacterium liquid to be measured after pre-treatment, extract DNA and carry out EMA-fluorescence quantitative PCR detection, calculated the viable bacteria concentration of this bacterium liquid to be measured by typical curve.Wherein, the process of this EMA-fluorescence quantitative PCR detection repeats no more, its reaction system and reaction conditions as follows:
Rt-PCR reaction system:
Note: Pb is 5S rRNA, and concentration is 2umol/L, and upstream primer F concentration is 9umol/L, and downstream primer R concentration is 9umol/L.Mix is TAKARA, 2x (DRR03P) and ROX(ROX II)
Rt-PCR reaction conditions:
3, experimental result and analysis:
Table 1
As can be seen from Table 1: slat chain conveyor counting process to the detected result of viable bacteria and EMA-fluorescence quantitative PCR method comparatively close to the detected result of viable bacteria, result explicit declaration: preprocessing process bacterium liquid to be measured carried out by pretreatment unit of the present invention, effectively achieve the DNA of dead bacterium in degraded bacterium liquid to be measured, avoid the generation of " false positive " result, ensure that the accuracy of detected result.
Table 2
| ? | EMA-quantitative fluorescent PCR (CFU/mL) viable count |
| 1st group | 1.27×10 8 |
| 2nd group | 1.20×10 8 |
| 3rd group | 1.15×10 8 |
As can be seen from Table 2: by EMA-fluorescence quantitative PCR method, parallel to carry out the result of living stems to three groups of bacterium liquid to be measured after pre-treatment is centrifugal all comparatively close, this absolutely proves: the pre-treatment carried out bacterium liquid to be measured by pretreatment unit of the present invention, effectively can ensure the stability of detected result.
Last it is noted that above each embodiment is only in order to illustrate technical scheme of the present invention, be not intended to limit; Although with reference to foregoing embodiments to invention has been detailed description, those of ordinary skill in the art is to be understood that: it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein some or all of technical characteristic; And these amendments or replacement, do not make the essence of appropriate technical solution depart from the scope of various embodiments of the present invention technical scheme.
Claims (11)
1. a living stems pretreatment unit, is characterized in that, described living stems pretreatment unit utilizes reagent E MA or PMA to implement the pretreatment unit of process to testing sample, comprising:
Housing;
Be arranged at the illumination system in this housing, refrigeration system and sample block;
Described illumination system is arranged at the top in described pretreatment unit housing, and described illumination system comprises light source, radiator fan and scatterer;
Described refrigeration system is arranged at the bottom in described pretreatment unit housing, and described refrigeration system comprises semiconductor cooler, radiator fan and scatterer;
Described sample block is arranged between described light source and semiconductor cooler, and the minimum perpendicular distance of described sample block and described light source is 2-10cm;
Wherein, described sample block is provided with the sample tube seat matched with sample hose, and this sample tube seat can make inserted sample hose have towards the tube wall face of light source, the axis of sample hose described in it and horizontal direction are that 10-70 ° of overturning angle is placed;
The top of described light source sets gradually scatterer and radiator fan composition illumination system, and described light source and described scatterer are formed and press laminating; Set gradually scatterer and radiator fan composition refrigeration system below described semiconductor cooler, and described scatterer and described semiconductor cooler form and press laminating;
Described sample block is positioned in refrigeration system, is formed press laminating by heat conductive silica gel fat and described semiconductor cooler, and the upper surface of the scatterer of refrigeration system and this semiconductor cooler are formed by heat conductive silica gel fat and press laminating;
Described light source is LED.
2. pretreatment unit according to claim 1, is characterized in that, described sample block is provided with multiple sample tube seat.
3. pretreatment unit according to claim 1, is characterized in that, described sample block is provided with the illuminance transducer monitoring light illumination, and the temperature sensor of energy monitoring system cooling system.
4. pretreatment unit according to claim 1, is characterized in that, the housing of this pretreatment unit is the tubular shell with upper shed, and has the lid that can fasten this housing seal, and described illumination system is arranged on this lid.
5. pretreatment unit according to claim 1, is characterized in that, the housing sidewall of this pretreatment unit is provided with opening, and be provided with can to the engaging support mechanism of this housing seal, and described sample block can take out through described opening or insert.
6. pretreatment unit according to claim 5, is characterized in that, described sample block is arranged in inner walls by rail structure or between the radiator fan of refrigeration system of housing bottom and scatterer.
7. the pretreatment unit according to any one of claim 1-6, is characterized in that, described housing is engineering plastics housing, and inner walls is mirror face stainless steel inwall.
8. pretreatment unit according to claim 1, is characterized in that, described sample block is aluminium alloy module.
9. pretreatment unit according to claim 1, is characterized in that, this pretreatment unit also comprises control mainboard, the supply unit of controlled light system and refrigeration system, and the display unit of the illumination that can show in preprocessing process and/or state of temperature.
10. a pretreatment process for living stems, is characterized in that, adopts the pretreatment unit as described in any one of claim 1-9, and the dead bacterium in testing sample is degraded, and described pretreatment process comprises the following steps:
The sample hose that testing sample and reagent are housed is placed in the sample tube seat that described pretreatment unit matches with sample hose;
Under light source, sequentially exposing irradiates sample hose, and Quality control module temperature is 2-8 DEG C, realizes the DNA of dead bacterium in degraded testing sample;
Wherein, described reagent comprises EMA or PMA.
11. 1 kinds are detected method of microorganism, it is characterized in that, comprise the following steps:
According to pretreatment process described in claim 10, pre-treatment is implemented to testing sample, the dead bacterium in testing sample is degraded;
Take out sample hose, DNA separation is carried out to this pretreated testing sample, and implements microorganism detection, and do not comprise medical diagnosis on disease and/or methods for the treatment of.
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| JP6804769B2 (en) * | 2015-12-28 | 2020-12-23 | 合同会社日本理化学開発 | A device for determining the state of living and dead bacteria, and a method for determining the state of living and dead bacteria using the device. |
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| CN202626164U (en) * | 2012-05-21 | 2012-12-26 | 河南省兽药监察所 | Device for performing covalent cross-linking on propylene glycol monomethyl acetate (PMA)/ ethyl methacrylate (EMA) and deoxyribonucleic acid (DNA) molecules of dead bacterial cell |
| CN203144414U (en) * | 2013-03-27 | 2013-08-21 | 北京博瑞立安生物技术有限公司 | Viable organism detection pre-treatment device |
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| DK1918385T3 (en) * | 2005-07-21 | 2011-09-19 | Morinaga Milk Industry Co Ltd | Microorganism detection method and microorganism detection kit |
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| CN101978275A (en) * | 2008-02-05 | 2011-02-16 | 普凯尔德诊断技术有限公司 | System for conducting the identification of bacteria in biological samples |
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Inventor after: Zhou Xu Inventor after: Dong Haiyan Inventor after: Chen Weidong Inventor after: Xie Ming Inventor after: Tommy Chou Inventor after: Li Xiaochen Inventor before: Zhou Xu Inventor before: Dong Haiyan Inventor before: Chen Weidong Inventor before: Xie Ming Inventor before: Tommy Chou Inventor before: Liu Xiaochen |
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Free format text: CORRECT: INVENTOR; FROM: ZHOU XU DONG HAIYAN CHEN WEIDONG XIE MING TOMMY CHOY LIU XIAOCHEN TO: ZHOUXU DONG HAIYAN CHEN WEIDONG XIE MING TOMMY CHOY LI XIAOCHEN |
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| C53 | Correction of patent for invention or patent application | ||
| CB03 | Change of inventor or designer information |
Inventor after: Zhou Xu Inventor after: Dong Haiyan Inventor after: Chen Weidong Inventor after: Tommy Chou Inventor after: Li Xiaochen Inventor before: Zhou Xu Inventor before: Dong Haiyan Inventor before: Chen Weidong Inventor before: Xie Ming Inventor before: Tommy Chou Inventor before: Li Xiaochen |
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| COR | Change of bibliographic data |
Free format text: CORRECT: INVENTOR; FROM: ZHOU XU DONG HAIYAN CHEN WEIDONG XIE MING TOMMY CHOY LI XIAOCHEN TO: ZHOU XU DONG HAIYAN CHEN WEIDONG TOMMY CHOY LI XIAOCHEN |
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